Antioxidant Activity of Hyptis suaveolens Poit.

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Life International Journal of Pharmacology ISSN 1811-7775 Life science alert ansinet Asian Network for Scientific Information

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International Journal of Pharmacology 4 (3): 227-229, 2008 ISSN 1811-7775 2008 Asian Network for Scientific Information Antioxidant Activity of Hyptis suaveolens Poit. U. Gavani and P.M. Paarakh Department of Pharmacognosy, The Oxford College of Pharmacy, Bangalore 560078, Karnataka, India Abstract: The antioxidant activity of the methanol extract of leaves of Hyptis suaveolens Poit. was evaluated in vitro by 1, 1-diphenyl-2-picrylhydrazyl (DPPII) radical scavenging activity using gallic acid and butylated hydroxyanisole (BHA) as reference standards. They exhibited strong antioxidant radical scavenging activity with IC, value of 0.4, 1.15 and 14.04 µg ml. ' for Gallic acid, BHA and Hyptis suaveolens, respectively. The antioxidant activity of methanol extract could be due to the presence of Flavonoids. Key words: Hyptis suaveolens Poit., antioxidant activity, DPPH, methanol extract, gallic acid, butylated hydroxyanisole INTRODUCTION Antioxidants are the substances used by the body to protect itself from the damage caused by oxidation. Oxidation is a chemical reaction that transfers electrons from a substance to an oxidizing agent. Oxidation reactions can produce free radicals, which starts chain reactions that damage cells. Free radical production is actually a normal part of life, part of the equation of simply breathing in oxygen. The body can cope with some free radicals and need them to function effectively. However, an overload of free radicals has been linked to many diseases, few of which include heart diseases (Aviram. 2000), liver diseases and cancers (Owen et al., 2000). The reactive oxygen species and free radicals produced in the cells include hydrogen peroxide, hypochlorous acid, hydroxyl radical and superoxide anion (Valko et al., 2007), which is known to cause cell damage by starting chemical chain reactions such as lipid peroxidation, or by oxidizing DNA or proteins (Sies, 1997). Hence the research has focused on use of antioxidants, with particular emphasis on naturally derived antioxidants, which may inhibit reactive oxygen species and may display protective effects. Plant phenolics, in particular phenolic acids, tannins and flavonoids are known to be potent antioxidants and occur in vegetables, fruits, nuts, seeds. roots and barks (Pratt and Hudson, 1990). In addition to their antioxidant properties, these compounds display a vast variety of pharmacological activities such as antiinflammatory, anticancer, alicarcinogenic aril antibacterial activities. Hyptis suaveolens Poit. (Laminaceae), known as Ganga tulsi, is an aromatic strongly scented herb found in Deccan Peninsula, North East India, Andaman and Nicobar Island, Philippines and Tropical America. In the Traditional System of Medicine, the plant is used as a stimulant, carminative for wounds, sudorific. galactogogue, catarrhal condition, infection of uterus, parasitic skin diseases (Anonymous, 2001). The phytoconstitutents isolated from the plant are hyptadienic acid (Raja et al, 1990), suaveolic acid suaveolol, methyl suaveolate, SS-sitosterol, olcanolic acid, ursolic acid, rosamarinic acid (Manchard et al., 1974), dehydroabietinol (Ziegler et al., 2002), 3SS-hydroxy lup-12-en-28-oic acid (Misra et al., 1983a), 3SS-hydroxyllup-20(29)-en-27-oic acid (Misra et al., 1983b) and essential oil (Peerzada, 1997). It has been reported to possess antibacterial (Asekum, 1999), wound healing (Shirwaikar et al., 2003), antiinflammatory activity (Grussi et al., 2006). antiplasmodial activity (Chukwujekwu et al., 2005). antimalarial (Ziegler et al., 2002) and antinociceptive (Santos et al., 2007) activities. Given the fact of traditional knowledge and the recent pharmacological studies, the Him in the present study was to evaluate is in vitro antioxidant DPPII free radical scavenging activity. MATERIALS AND METHODS Chemicals: 1,1-diphenyl-2-picrylhydrazyl (DPPH), gallic acid and butylated hydroxyanisole was purchased from Loba Chemie Pvt. Ltd., Mumbai. All the chemicals and reagents used were of analytical grade. Corresponding Author: Dr. P.M. Paarakh, Department of Pharmacognosy. The Oxford College of Pharmacy, J.P. Nagar, I. Phase. Bangalore 560078, Karnataka, India Tel: 09880681532 Fax: 080-26548658 227

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Int. J. Pharmacol., 4 (3): 227-229, 2008 Plant material: Fresh leaves of Hyptis suaveolens Poit. were collected from Bangalore, Kamataka in July 2007. shade dried and authenticated by Prof. Gajendra Rao, Central Council for Research in Ayurveda and Siddha. Bangalore. Voucher specimen is deposited in the Department of Pharmacognosy. The Oxford College of Pharmacy, Bangalore. Preparation of methanol extract: The shade dried. powdered leaves (450 g) were exhaustively extracted with petroleum ether (60-80°C) followed by chloroform and methanol using soxhlet apparalus. The methanol extract. (HSME) was concentrated in vacuum (yield: 1.86% w/w) and was only used for antioxidant activity. Phytochemical screening: The coarse powder of leaves of Ilyptis suaveolens (25 g) was subjected to successive extraction with different solvents in their increasing order of polarity from petroleum ether (60-80°C), chloroform. methanol and water. The extracts were concentrated and subjected to various chemical tests to detect the presence of different phytoconstitutents (Khandelwal, 2003). Evaluation of antioxidant activity: The free radical scavenging activity of the methanol extract of Hyplis suaveolens was measured by 1,1-diphenyl-2pierylhydrazyl (DPPH) using the methox described by Shimoda et al. (1992). 0.1 mM solution of DPPII in ethanol was prepared. One milliliter of the solution was added to 3 mL of IISME solution in methanol at different concentration (100-1.56 ug mL). The mixture was shaken vigorously and allowed to stand at room temperature for 30 min. Then the absorbance was measured at 517 nm by using a spectrophotometer (UV-VIS Shimadzu). Reference standard compound being used was gallic acid and butylated hydroxyanisole 0.125-1 and 1-5 μg mL, respectively. The ICs, value is the concentration of sample required to inhibit 50% of the DPPH free radical. The IC value for the sample was calculated using log dose inhibition curve. Lower absorbance of the reaction mixture Table 1: Phytochemical screening of different extracts indicated higher free radical activity. The percent DPPH scavenging effect was calculated using the following equation: DPPH scavenging effect (%) = 100×A/A, where, A, was the absorbance of the control reaction and A was the absorbance in presence of the standard sample or IISME. RESULTS AND DISCUSSION Preliminary phytochemical screening: Preliminary phytochemical screening revealed the presence of steroids, flavonoids, alkaloids, carbohydrates and tannins in the different extracts (Table 1). From the literature, it is clear that phytoconstituents viz., steroids only have been isolated from different extracts, still other phytoconstituents are yet to be isolated. Only methanol extract was chosen as it contains flavonoids which are generally potent inhibitors of free radicals (Havsteen, 1983). DPPH radical scavenging activity: As shown in Table 2. HSME has polent. DPPH radical scavenging activity with an IC value of 14.04 µg mL¯¯. IC value of gallic acid and butylated hydroxyanisole were found to be 0.4 and 1.15 μg mL, respectively. DPPH is a stable free radical that can accept an electron or hydrogen radical to become a stable diamagnetic molecule. DPPH radicals react with suitable reducing agents then losing colour sticchiometrically with the mumber of electrons consumed which is measured spectrometrically at 517 nm (Soares et al., 1997; Lugasi et al., 1998). Gallic acid is a potent. free radical scavenging and butylated hydroxyanisole is known antioxidant and is used as preservative. So when compared to such pure components, IC, value of crude HSME is quite good proving that it is potent DPPH free radical scavenger. This can be attributed to flavonoids. Extracts Petroleum ether Chloroform Methanol Aqueous (water) Presence; Absent Steroids Alkaloids Glycosides Saponin Flavonoid Tannin Carbohydrates Table 2: DPPH free radical scavenging activity of methanol extract of Hypfis suaveolens [HSME] Extract/standard Gallic acid BHA HSME IC value (ug ml. '; mean1ST.M) 0.40±2.68 1.15±3.56 14.04±2.08 228

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